Defining HIV Env protein expression in latently infected cells

定义潜伏感染细胞中的 HIV 包膜蛋白表达

基本信息

批准号：
10762524
负责人：
Alberto Bosque
金额：
$ 24.23万
依托单位：
GEORGE WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-08-04 至 2025-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10762524
关键词：
Address Affect Antibodies B-Lymphocytes Binding Biological Assay Cell Separation Cell model Cell surface Cells Cellular biology Characteristics Chronic Clinical Trials Data Detection Development Effector Cell Epitopes Flow Cytometry Future Gender Genetic Genetic Variation HIV HIV Core Protein p24 HIV-1 Immune Immunotherapy In Vitro Infection Interruption Investigation Knowledge Lead Learning Length Measures Mediating Methodology Methods Modeling Molecular Conformation Molecular Virology Outcome Persons Phagocytosis Protein Conformation Proteins Publishing RNA RNA Splicing Role Surface Testing Therapeutic Therapeutic Uses Transcript Translating Translations Viral Viral Proteins Virus Virus Replication antibody-dependent cell cytotoxicity antiretroviral therapy cohort env Gene Products flexibility gag Gene Products high reward high risk improved lonely individuals monomer neutralizing antibody novel prevent protein expression reactivation from latency treatment strategy virus genetics

Address Affect Antibodies B-Lymphocytes Binding Biological Assay Cell Separation Cell model Cell surface Cells Cellular biology Characteristics Chronic Clinical Trials Data Detection Development Effector Cell Epitopes Flow Cytometry Future Gender Genetic Genetic Variation HIV HIV Core Protein p24 HIV-1 Immune Immunotherapy In Vitro Infection Interruption Investigation Knowledge Lead Learning Length Measures Mediating Methodology Methods Modeling Molecular Conformation Molecular Virology Outcome Persons Phagocytosis Protein Conformation Proteins Publishing RNA RNA Splicing Role Surface Testing Therapeutic Therapeutic Uses Transcript Translating Translations Viral Viral Proteins Virus Virus Replication antibody-dependent cell cytotoxicity antiretroviral therapy cohort env Gene Products flexibility gag Gene Products high reward high risk improved lonely individuals monomer neutralizing antibody novel prevent protein expression reactivation from latency treatment strategy virus genetics

展开

项目摘要

Project Summary/Abstract Advances in B cell biology and molecular virology have enabled the discovery, characterization, and commercial development of broadly neutralizing antibodies (bNAbs). They are a promising immunotherapy that can be incorporated in many strategies for the treatment and/or cure of HIV-1. Antibodies have many effector functions beyond neutralization and their most important mechanism of action for treatment/cure may be their ability to opsonize infected cells, tagging them for antibody-dependent cellular cytotoxicity or phagocytosis (ADCC/ADCP) by immune effector cells. It is not clear that bNAbs are mediating clearance of infected cells in clinical trials when they are passively infused into chronically infected people living with HIV (PLWH). An obvious reason for this lack of efficacy of bNAbs may be a lack of Env protein expression while on suppressive ART. Our lack of knowledge about the level of Env expression during HIV latency and latency reversal prevents effective use of bNAbs as therapeutics. To address this gap, the ability to detect low level Env protein expression in cells infected with different subtypes of HIV-1 is needed. The Bosque lab has published an ultrasensitive method to detect p24 Gag protein down to the fg/ml level and has preliminary data for a newly developed Env assay. Here we propose to use these assays to detect low levels of Env and Gag protein in a well-described model of latency and compare the data to Env surface expression as measured by flow cytometry (Aim 1). Within these assays, we have the flexibility to test Env detection using bNAbs targeting different epitopes to probe for Env conformation (i.e., trimer, monomer, etc) and extend these analyses to diverse HIV isolates from different subtypes to measure differences in Env expression and latency attributable to genetically diverse virus (Aim 2). Knowing these crucial factors about Env and Gag expression in a carefully controlled model of latency will allow us to further investigate HIV protein translation in future studies using cohorts of PLWH. These studies will improve the ability of therapeutics using Env-targeting strategies to target latently infected cells.

项目摘要/摘要 B细胞生物学和分子病毒学的进步已使发现，表征和商业开发广泛中和抗体（BNAB）。它们是一种有希望的免疫疗法将HIV-1治疗和/或治疗的许多策略纳入其中。抗体具有许多效应子功能除了中和及其最重要的治疗/治疗作用机制可能是他们的能力对感染的细胞进行调整，将其标记为抗体依赖性细胞细胞毒性或吞噬作用（ADCC/ADCP）通过免疫效应细胞。目前尚不清楚BNAB在临床试验中介导感染细胞的清除率他们被动地注入了感染艾滋病毒（PLWH）的长期感染者。一个明显的原因在抑制艺术上，BNAB缺乏疗效可能是缺乏Env蛋白的表达。我们缺乏关于在艾滋病毒潜伏期和潜伏期逆转过程中的ENV表达水平的知识可有效使用 bnabs作为治疗学。为了解决这一差距，检测感染细胞中低水平的Env蛋白表达的能力需要使用不同的HIV-1亚型。 Bosque Lab已发表了一种超敏化方法来检测P24 GAG蛋白降至FG/ML水平，并具有新开发的ENV分析的初步数据。我们在这里提出使用这些测定法以延迟的描述模型检测低水平的ENV和GAG蛋白通过流式细胞仪测量的数据表达表达的数据（AIM 1）。在这些测定中，我们有使用针对不同表位的BNABS测试ENV检测的灵活性，以探测ENV构象（即Trimer，，单体等）并将这些分析扩展到来自不同亚型的不同HIV分离株以测量差异在ENV表达和延迟归因于遗传多样的病毒（AIM 2）。知道这些关键因素关于在经过精心控制的延迟模型中的Env和GAG表达，将使我们能够进一步研究HIV 在未来研究中使用PLWH的蛋白质翻译。这些研究将提高治疗能力的能力使用靶向靶向策略来靶向潜在感染的细胞。